The present invention relates to methods of reducing hyperlipidemia and hyperglycemia, as well as to certain compounds and compositions.
The adverse complications of hyperlipidemia are prevalent worldwide, manifest by the high incidence of serious atherosclerotic disease including heart attack and stroke. Cardiovascular and cerebrovascular disease are slowly progressive conditions, often undetected in their early stages of pathophysiology, that are typified by elevated circulating levels of lipids and demonstrable by the accumulating deposition of fatty substances in the macrovasculature over years to decades. Only when sufficient vascular damage and accumulated thrombotic material is present to occlude a major vessel or become detached and create a vascular obstruction, does this disease wreak its high incidence of morbidity and mortality.
Underlying factors both within and beyond an individual""s control contribute to the disease process, including genetic predisposition, co-morbidity with diseases such as diabetes mellitus, or lifestyle factors including diet, smoking, and exercise, among other factors. While genetic components have only recently become potential direct targets for therapeutic intervention, and alterations in lifestyle are difficult to achieve and often difficult to maintain, therapies directed toward pharmacologically lowering circulating lipid levels have been clinically successful at reducing the incidence of cardiovascular and cerebrovascular disease. Elevated circulating lipids and their markers indicative of a pathological state include free fatty acids, glycerol, triglycerides, cholesterol, and low-density lipoprotein. Reduction or control of each of these components can have a positive impact upon other abnormal circulating analytes, including glucose and insulin.
Diabetes mellitus is a disease of multifactorial origins involving disorders of metabolism associated with insulin-producing as well as insulin-regulated tissues. In addition to being a disease of hyperglycemia, some forms of the disease are associated with abnormally elevated circulating lipid levels, Therefore, lowering the circulating levels of lipids and glucose is beneficial for the treatment of patients with diabetes mellitus. In type II diabetes, while genetic mutations can cause certain subsets of the disease (e.g., maturity-onset diabetes of the young, or MODY), the most common form of the disease involves resistance to the action of insulin on insulin-target tissues, and is associated with obesity, predominately of abdominal origin. In addition to elevated glucose levels, circulating levels of triglycerides, cholesterol and non-esterified fatty acids (NEFA) are elevated during a fasting period. Persons afflicted with diabetes are predisposed to the development of cardiovascular disorders, which is the main cause of morbidity and mortality for this disease. Resistance to insulin action occurs not only in hyperglycemic states but also in Syndrome X, an insulin-resistant disorder characterized by hyperinsulinemia and dyslipidemia. (Reaven GM, Syndrome X, Clinical Diabetes, March/April 1994:32-36). In this state, normoglycemia can be maintained due to an increase in the secretion of insulin. However, when the insulin-resistance cannot be sustained by the compensatory activity of the pancreas, glucose-intolerance supervenes. Thus, lipid abnormalities, coupled either with overt hyperglycemia or with the normoglycemic Syndrome X phenotype, are believed to contribute to the severity of diabetic complications, such as cardiovascular complications (Olefsky, JM, Current Approaches to the Management of Type 2 Diabetes: A Practical Monograph, National Diabetes Education Initiative, 1997).
Oral antihyperglycemic agents, such as the sulfonylureas and biguanides, promote improvements in elevated lipid levels through overcoming the associated state of insulin-resistance, either by potentiating the endogenous release of insulin from the xcex2 cells of the pancreas, as in the case of the sulphonylureas, or by enhancing glucose disposal and reducing gluconeogenesis, as in the case of biguanides. Other approaches to controlling the elevated lipids and glucose have involved the use of broad acting oral antihyperglycemics such as the thiazolidinedione class of agents which lower fatty acids and improve the insulin-resistant state of insulin responsive tissues. The compound 4,7,8xcex1H-eudesma-5(6),11(13)-dien-8,12-olide, also known as helenin and alantolactone, has been reported to induce hyperglycemia in rabbits at high dose, to induce hypoglycemia at a moderate dose, and to inhibit hyperglycemia induced by food (1986, Handbook of Effective Ingredients of Medicinal Plants, Beijing, China).
Numerous, diverse therapeutic strategies have been developed for treating the hyperlipidemia and associated hyperglycemia of type II diabetes. The aim of lowering of circulating lipids has been to reduce the cardiovascular morbidity and/or improving the overall diabetic state, Examples of classes of agents acting directly on plasma triglyceride and cholesterol content include the HMG-CoA reductase inhibitors, fibric acids, and bile-salt resins. Whereas these classes are effective in lowering triglyceride and cholesterol content, they have little impact on plasma fatty acids.
Of the lipid classes, non-esterified fatty acids appear to play a role in promoting the diabetic and/or the hyperlipidemic, insulin-resistant, state. Elevated free fatty acids arise from either the excess body burden of adipose tissue in the obese state or from uncontrolled breakdown of triglycerides in adipose tissue, a major insulin-target tissue, or both. In addition, elevated levels of free fatty acids have been shown to acutely induce insulin resistance in muscle, the major glucose utilizing tissue of the body, where a direct effect on glucose transport in muscle is observed. In addition, fatty acids affect liver metabolism to increase hepatic glucose output. Furthermore, elevated fatty acids induce impaired xcex1-cell functioning by lessening the secretion of insulin from xcex1-cells in response to a glucose stimulus. Attempts to reduce fatty acid levels have included the development of thermogenic agents (?3 agonists) which are believed to function by stimulating brown adipose tissue to oxidize fatty acids and thus clear them from the circulation. Alternatively, removing fatty acids from their influence in the liver was attempted by developing inhibitors of fatty acid oxidation by the enzyme carnitine palmitoyl transferase I (CPTI).
In particular, resistance to the action of insulin in adipose tissue appears to plays a significant role in the elevation of fatty acids. Agents that inhibit adipose tissue breakdown of triglycerides are known as lipolytic inhibitors. It is toward the development of new methods and agents for lowering circulating lipids to treat hyperlipidemic disorders or for lowering circulatory glucose to treat hyperglycemia, diabetes and associated disorders that the present invention is directed.
The invention relates to lipid-lowering or glucose-lowering methods and compounds, particularly lipid-lowering or glucose-lowering compounds, with an A-B ring structure of the following Formula I: 
wherein the dotted line between carbons 5 and 6, the dotted line between carbon 4 and R2, and the dotted line between carbon 11 and R3 each independently represent an optional double bond from the respective numbered carbon to a linked carbon.
The present invention to provides, among other things: methods for lowering lipid levels in a mammal to treat or prevent the development of pathology associated with elevated lipid levels; compounds which inhibit the enzyme hormone-sensitive lipase and inhibit lipolysis; agents and methods for the treatment of insulin resistance and Syndrome X; methods for lowering glucose levels in a mammal in order to treat or prevent the development of pathology associated with hyperglycemia; methods for treating impaired glucose tolerance associated with fasting or prandial hyperglycemia, normoglycemia, or insulin resistance, for example to delay or prevent the onset of non-insulin-dependent diabetes mellitus.
In accordance with the present invention, methods for lowering lipid levels in a mammal or lowering glucose levels in a mammal (particularly a hyperglycemic mammal) are provided comprising administering to said mammal an effective amount of a composition comprising a compound of the formula:
The invention relates to a method for lowering blood lipid levels or lowering blood glucose in a mammal comprising administering to the mammal a lipid-lowering or glucose-lowering effective amount of one or more compounds with an A-B ring structure, the compounds of Formula I, wherein the dotted line between carbons 5 and 6, the dotted line between carbon 4 and R2, and the dotted line between carbon 11 and R3 each independently represent an optional double bond from the respective numbered carbon to a linked carbon, wherein R1 is hydrogen, hydroxy, a lower alkanoyloxy or an aroyloxy group (wherein aryl of aroyloxy can be substituted by one or more lower alkyl, lower alkoxy, halo, trifluoromethyl, di(lower)alkylamino, hydroxy, nitro or C1-C3 alkylenedioxy groups); wherein X is hydroxy or, together with Y is oxy linking carbon 8 and carbon 12; wherein Y is hydroxy or an amino group that can be mono or di-substituted with lower alkyl, where the lower alkyls can be joined to form a five or six-membered ring; wherein R2 and R3 are each independently a lower alkyl or alkenyl group, wherein the lower alkyl of R3 can be substituted at the carbon linked to C11 with an amino group that can be mono or di-substituted with lower alkyl, where the lower alkyls can be joined to form a five or six-membered ring, or pharmaceutically acceptable salts thereof, with the proviso that where R1is hydrogen, either (a) X is hydroxy or (b) R3 is substituted with amino. The invention further relates to the administered pharmaceutical composition.
The invention also relates to a compound (and pharmaceutical compositions thereof) of Formula I, wherein the dotted line between carbons 5 and 6, the dotted line between carbons 4 and R2, and the dotted line between carbon 11 and R3 each independently represent an optional double bond from the respective numbered carbon to a linked carbon wherein R1 is hydrogen, hydroxy, a lower alkanoyloxy or an aroyloxy group (wherein aryl of aroyloxy can be substituted by one or more lower alkyl, lower alkoxy, halo, trifluoromethyl, di(lower)alkylamino, hydroxy, nitro or C1-C3 alkylenedioxy groups); wherein X is hydroxy or, together with Y is oxy linking carbon 8 and carbon 12; wherein Y is hydroxy or an amino group that can be mono or di-substituted with lower alkyl, where the lower alkyls can be joined to form a five or six-membered ring; wherein R2 and R3 are each independently a lower alkyl or alkenyl group, wherein the lower alkyl of R3 can be substituted with an amino group that can be mono or di-substituted with lower alkyl, where the lower alkyls can be joined to form a five or six-membered ring, or pharmaceutically acceptable salts thereof, with the provisos that (1) where X is hydroxy, then R1 is not hydrogen, (2) where R1 is hydroxy, then X is either (a) X is hydroxy or (b) R3 is substituted with amino, and (3) where R2 is methylene linked to C4 by a double bond, R3 is methylene linked to C11 by a double bond, and X is oxy, then R1 is not ethanoyloxy. Preferably, R1is not propanoyloxy.
In one embodiment, the hydrogen substituents at positions 5, 7 and 8 are alpha relative to the A-B ring structure. In another embodiment, X is hydroxy. In another embodiment, R1 is a lower alkanoyloxy group, for example, selected from the group consisting of acetoxy, propanoyloxy, isobutyryloxy, and cyclopropanoyloxy. The compound is, for example, selected from the group consisting of 2xcex1-acetoxy-5,7,8xcex1H-eudesma-4(15),11(13 )-dien-8,12-olide; 2xcex1-propanoyloxy-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide; 2xcex1-cyclopropanoyloxy-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide; 2xcex1-acetoxy-4,5,7,8,11xcex1H-eudesman-8,12-olide; 2xcex1-cyclopropanoyloxy-5,7,8,11xcex1H-eudesm-4(15)-en-8,12-olide. In another embodiment, R1 is a aroyloxy group, such as furoyloxy and benzoyloxy, for example, 2xcex1-benzoyloxy -5,7,8xcex1H-eudesma-4(15),11(13 )-dien-8,12-olide; 2xcex1-furoyloxy-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide, 2xcex1-benzoyloxy-4,5,7,8,11xcex1H-eudesman-8,12-olide; or 2xcex1-benzoyloxy-5,7,8,11xcex1H-eudesm-4(15)-en-8,12-olide.
The compounds can be administered in a lipid-lowering effective amount or a a glucose-lowering effective amount.
In one embodimemt, (1) X is oxy, (2) R1 is lower alkanoyloxy or aroyloxy, with the proviso that when R2 is methylene linked to C4 by a double bond, R3 is methylene linked to C11 by a double bond, then R1 is not ethanoyl, and (3) wherein R2 and R3 are each independently a lower alkyl group or a lower alkenyl group with the unsaturation limited to the linkage to carbon 4 or carbon 11. In another embodiment, (1) X is oxy, (2) R1 is lower alkanoyloxy or aroyloxy, and (3) wherein R2 and R3 are each independently a lower alkyl group or a lower alkenyl group with the unsaturation limited to the linkage to carbon 4 or carbon 11.
The lower alkyl and alkenyl groups contain one to six carbons. Lower alkenyl groups are defined in conjunction with the backbone structures of Formula I, such that a methylene group has its unsaturation in conjunction with, for example, carbon 4 or carbon 11. In one embodiment, at least one or both of R1 and R2 are methyl or methylene (both bonds in a double bond to carbon 4 or carbon 11). The lower alkanoyloxy groups referred to above contain one or two to six carbon atoms and include methanoyloxy, ethanoyloxy or acetoxy, propanoyloxy, butyryloxy, pentanoyloxy, hexanoyloxy, and the corresponding branched-chain isomers thereof Also included among the alkanoyloxy groups herein are those comprising substituted and unsubstituted cycloalkyl groups, such as cyclopropanoyloxy, cyclobutanoyloxy, cyclopentanoyloxy, cyclohexanoyloxy, 1-methylcyclohexanoyloxy, trans-4-isopropylcyclohexanoyloxy, and the like. The aroyloxy groups referred to herein include substituted and unsubstituted aromatic and heteroaromatic groups such as furoyloxy, benzoyloxy, 4-fluorobenzoyloxy, 2-chlorobenzoyloxy, 2-methoxybenzoyloxy, 3-ethylbenzoyloxy, 4-ethylbenzoyloxy, 4-isopropylbenzoyloxy, 4-chlorobenzoyloxy, 3,5-tert-dibutyl-4-hydroxybenzoyloxy, 2-methoxy-5-chlorobenzoyloxy, 3-fluoro-4xe2x80x2-methoxybenzoyloxy, 2-trifluoromethylbenzoyloxy, 4-trifluoromethylbenzoyloxy, 2-naphthoyloxy, 2-thiophenecarbonyloxy, 3-pyridinecarbonyloxy, and 5-methyl-2xe2x80x2-pyrazinecarbonyloxy. Both the 2xcex1 and 2xcex2 isomers are included, 2xcex1 is preferred. The lower alkyl groups include methyl, ethyl, propyl, butyl, pentyl, heyxl, and the corresponding brnached-chain isomers thereof Preferably, the aryl groups are C6-C10 aromatic groups, or a heteroaromatic groups with 5 to 10 ring atoms, of which, preferably, up to two (2) are heteroatoms selected from nitrogen, oxygen or sulfur.
The lower alkyl groups referred to above include methyl, ethyl, propyl, butyl, pentyl, hexyl, and the corresponding branched-chain isomers thereof As described above, the bond linking groups R2 and R3 can be a single or double bond, and the alkyl groups described herein can be bonded to carbons 4 and 11, respectively, by means of a single or double bond.
Non-limiting examples of compounds in which R1 is hydrogen include salts, such as sodium salts, of: 8xcex2-hydroxy-4,7,8xcex1H-eudesama-5,11(13)-dien-12-oic acid; 8,xcex2-hydroxy-5,7,8,11xcex1H-eudesam-4(15)-en-12-oic acid; 8xcex2-hydroxy-4,5,7,8,11xcex1H-eudesman-12-oic acid; 2xcex1-hydroxy-4(15),11(13)eudesmadien-12,8-olide. Non-limiting examples of compounds in which R1 is hydroxy include salts, such as sodium or hydrochloride salts, of: 2xcex1,8xcex2-dihydroxy-2xcex2-4,5,7,8,11xcex1H-eudesaman-12-oic acid; 2xcex1,8xcex2-dihydroxy-4(15)11(13)-eudesamadien-12-oic acid; 2xcex1-benzoyl-8xcex2-hydroxy-2xcex2, 4, 5, 7, 8, 11xcex1H-eudesaman-12-oic acid; 1-[2xcex1-hydroxy-11,12-dihydro-5,7,8xcex1H-eudesm-4(15)-en-8,12-olidyl]-pyrrolidine; and 1-[2xcex1-benzoyloxy- 11,12-dihydro-5,7,8xcex1H-eudesm4(15)-en-8,12-olidyl]-pyrrolidine.
Compounds in which R1 is a lower alkanoyloxy group include but are not limited to 2xcex1-acetoxy-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide; 2xcex1-propanoyloxy-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide; 2xcex1-isobutyryloxy-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide; xcex1-cyclopropanoyloxy-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide; 2xcex1-acetoxy-4,5,7,8,11xcex1H-eudesman-8,12-olide; and 2xcex1-cyclopropanoyloxy-5,7,8,11xcex1H-eudesma-4(15)-en-8,12-olide. Compounds in which R1 is a aroyloxy group include. for example, 2xcex1-benzoyloxy-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide,2xcex1-furoyloxy-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide; 2xcex1-benzoyloxy-4,5,7,8,11xcex1H-eudesman-8,12-olide; and 2xcex1-benzoyloxy-5,7,8,11xcex1H-eudesm-4(15)-en-8,12-olide.
The compounds of the present invention, upon administration to hyperlipidemic mammals, exact positive effects on circulating lipid levels. Furthermore, in vitro, the compounds specifically inhibit an enzyme, hormone-sensitive lipase (HSL), and inhibit lipolysis in a whole-cell model. In vivo, the compounds described herein inhibit the appearance of increased circulating free fatty acids in overnight fasted normal mice. In genetic models of obesity/type II diabetes associated with hyperlipidemia, the compounds reduce elevated circulating lipid levels.
The compounds of the present invention are prepared from synthetic starting materials or those isolated from natural sources. One particular starting material with double bonds between carbons 4 and 15, and between carbons 11 and 13,2xcex1-hydroxy-5,7,8xcex1H-eudesma4(15),11(13)-dien-8,12-olide, can be synthesized by the procedure described by Tomioka et al. (1984, Tetrahedron Letters 25:333-336) or isolated and purified from the plant Iva microcephala or other plants, as described by Herz et al. (1962, J Org. Chem. 27:905-910). The alkanoyloxy and aroyloxy derivatives of 2-hydroxy compounds, such as 2xcex1-hydroxy-5,7,8xcex1H-eudesma-4(15),11(13)dien-8,12-olide, are prepared by synthetic procedures known to one of ordinary skill; for example, 2xcex1-propanoyloxy-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide and 2xcex1-benzoyloxy-5,7,8xcex1H-eudesma-4(15),11(13)dien-8,12-olide are prepared from 2(xcex1-hydroxy-5,7,8xcex1H-eudesm-4(15),11(13)-dien-8,12-olide by reaction with propanecarbonyl chloride and benzoyl chloride, respectively. To prepare the compounds of the present invention that have single bonds between R2 and R3 and the respective ring carbon, a dien such as 2xcex1-hydroxy-5,7,8xcex1H-eudesm-4(15),11(13)-dien-8,12-olide is first reduced to 2xcex1-hydroxy-4,5,7,8,11xcex1H-eudesman-8,12-olide, for example following the procedure of Herz et al. (1962, J Org. Chem. 27:905-910), and then reacted with, for example, acyl chloride or benzoyl chloride, to yield 2xcex1-acetoxy-4,5,7,8,11xcex1H-eudesman-8,12-olide and 2xcex1-benzoyloxy-4,5,7,8,11xcex1H-eudesman-8,12-olide, respectively. Those compounds wherein only one of R2 or R3 is double-bonded to the ring can be isolated or prepared by synthesis or from natural sources, followed by synthetic procedures similar to those described above For example compounds 5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide, 4,7,8xcex1H-eudesma-5(6),11(13)-dien-8,12-olide and 5,7,8,11xcex1H-eudesm-4(15)-en-8,12-olide can be isolated from 80% isohelenin (which is available from Sigma Chemical Co., St. Louis, Mo., USA) by silica gel chromatography (e.g., loading with petroleum, ether, eluting with increasing concentrations of ethyl ether). 4,5,7,8,11xcex1H-eudesman-8,12-olide can be prepared by the reduction of 4,7,8xcex1H-eudesma-5(6),11(13)-dien-8,12-olide.
A useful source of compounds or starting materials are the plant isolates described by numerous authors including: Bohlmann et al., xe2x80x9cNew Sesquiterpene Lactones from Inula Species,xe2x80x9d 1978, Phytochem. 17:1165-1172, Topcu et al., xe2x80x9cCytotoxic and Antibacterial Sesquiterpenes from Imula Graveolens,xe2x80x9d 1993, Phytochem. 33:407-410, Ohta et al., xe2x80x9cSequiterpene Constituents fo Two Liverworts of Genus Diplophyllum, 1977, Tetrahedron 33:617-628; Herz et al., 1964, xe2x80x9cConstituents of Iva Species II,xe2x80x9d J Org. Chem. 29:1022-1026.
Compounds where R3 incorporates an amino can be synthesized by Michael addition, such as the analogous such additions described in Hejchman et al. (1995, 38:3407-3410). Open ring compounds can be synthesized by hydrolysis of the corresponding lactone.
Typical synthetic procedures are described in the Examples, below. Both the 2xcex1 and 2xcex2 isomers are included; 2xcex1 is preferred. The 2xcex2 isomer can be prepared from the corresponding 2xcex1 isomer using the Mitsunobu reaction (Mitsunobu et al., 1967, Bull. Chem. Soc. Japan 40:935).
Representative compounds of the present invention include:
2xcex1-cyclohexanecarbonyloxy-4,5,7,8,11xcex1H-eudesman-8,12-olide
2xcex1-cyclopentanecarbonyloxy-4,5,7,8,11xcex1H-eudesman-8,12-olide
2xcex1-cyclobutanecarbonyloxy-4,5,7,8,11xcex1H-eudesman-8,12-olide
2xcex1-(2xe2x80x2-naphthoyloxy)5,7,8,11xcex1H-eudesm-4(15)-en-8,12-olide
2xcex1-(1xe2x80x2-methylcyclohexanecarbonyloxy)-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide
2xcex1-(4xe2x80x2-trifluoromethylbenzoyloxy)-4,5,7,8,11 xcex1H-eudesman-8,12-olide
2xcex1-(2xe2x80x2-naphthoyloxy)-4,5,7,8,11xcex1H-eudesman-8,12-olide
2xcex1-cyclohexanecarbonyloxy-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide
2xcex1-(3xe2x80x2,5xe2x80x2-di-tert-butyl-4xe2x80x2-hydroxybenzoyloxy)5,7,8,11xcex1H-eudesm-4(15)-en-8,12-olide
2xcex1-(2xe2x80x2-methoxy-5xe2x80x2-chlorobenzoyloxy)-4,5,7,8,11xcex1H-eudesman-8,12-olide
2xcex1-(trans-4xe2x80x2-isopropylcyclohexanecarbonyloxy)-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide
2xcex1-cyclopentanecarbonyloxy-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide
2xcex1-(4xe2x80x2-ethylbenzoyloxy)5,7,8,11xcex1H-eudesm-4(15)-en-8,12-olide
2xcex1-(3xe2x80x2-ethylbenzoyloxy)5,7,8,11xcex1H-eudesm-4(15)-en-8,12-olide
2xcex1-cyclobutanecarbonyloxy-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide
2xcex1-(2xe2x80x2-methoxybenzoyloxy)5,7,8,11xcex1H-eudesm-4(15)-en-8,12-olide
2xcex1-(2xe2x80x2-trifluoromethylbenzoyloxy)-5,7,8,11xcex1H-eudesm-4(15)-en-8,12-olide
2xcex1-(3xe2x80x2-fluoro-4xe2x80x2-methoxybenzoyloxy)-5,7,8,11xcex1H-eudesm-4(15)-en-8,12-olide
2xcex1-(2xe2x80x2-methoxy-5xe2x80x2-chlorobenzoyloxy)5,7,8,11xcex1H-eudesm-4(15)-en-8,12-olide
2xcex1-(2xcex1-(2xe2x80x2-chlorobenzoyloxy)-5,7,8,11xcex1H-eudesm-4(15)-en-8,12-olide
2xcex1-(4xe2x80x2-fluorobenzoyloxy)-5,7,8,11xcex1H-eudesm4(15)-en-8,12-olide
2xcex1-(trans-4xe2x80x2-isopropylcyclohexanecarbonyloxy)-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide
2xcex1-(1xe2x80x2-methylcyclohexanecarbonyloxy)-5,7,8xcex1H-eudesma-4( 15),11(13)-dien-8,12-olide
2xcex1-(4-fluorobenzoyloxy)-4,5,7,8,11xcex1H-eudesman-8,12-olide
2xcex1-(2xcex1-(2xe2x80x2-chlorobenzoyloxy)-4,5,7,8,11xcex1H-eudesman-8,12-olide
2xcex1-(2xe2x80x2-methoxybenzoyloxy)-4,5,7,8,11xcex1H-eudesman-8,12-olide
2xcex1-(3xe2x80x2-ethylbenzoyloxy)-4,5,7,8,11xcex1H-eudesman-8,12-olide
2xcex1-(4xe2x80x2-ethylbenzoyloxy)-4,5,7,8,11xcex1H-eudesman-8,12-olide
2xcex1-(4xe2x80x2-isopropylbenzoyloxy)-4,5,7,8,11xcex1H-eudesman-8,12-olide
2xcex1-(4xe2x80x2-chlorobenzoyloxy)-4,5,7,8,11xcex1H-eudesman-8,12-olide
2xcex1-(3xe2x80x2,5xe2x80x2-tert-dibutyl-4xe2x80x2-hydroxybenzoyloxy)-4,5,7,8,11xcex1H-eudesman-8,12-olide
2xcex1-(2xe2x80x2-naphthoyloxy)-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide
2xcex1-(3xe2x80x2-fluoro-4xe2x80x2-methoxybenzoyloxy)-4,5,7,8,11xcex1H-eudesman-8,12-olide
2xcex1-(2xe2x80x2-trifluoromethylbenzoyloxy)-4,5,7,8,11xcex1H-eudesman-8,12-olide
2xcex1-(4xe2x80x2-isopropylbenzoyloxy)- 5,7,8,11xcex1H-eudesm-4(15)-en-8,12-olide
2xcex1-(4xe2x80x2-chlorobenzoyloxy)5,7,8,11xcex1H-eudesm-4(15)-en-8,12-olide
2xcex1-cyclohexanecarbonyloxy-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide
2xcex1-cyclopentanecarbonyloxy-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide
2xcex1-(4xe2x80x2-trifluoromethylbenzoyloxy)-5,7,8,11xcex1H-eudesm-4(15)-en-8,12-olide
2xcex1-cyclobutanecarbonyloxy-5,7,8xcex1H-eudesma-4(15). 11 (1 3)-dien-8,12-olide
2xcex1-(1xe2x80x2-methylcyclohexanecarbonyloxy)-4,5,7,8,11xcex1H-eudesman-8,12-olide
2xcex1-(trans-4xe2x80x2-isopropylcyclohexanecarbonyloxy)-4,5,7,8,11xcex1H-eudesman-8,12-olide
2xcex1-(4xe2x80x2-fluorobenzoyloxy)-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide
2xcex1-(2xcex1-(2xe2x80x2-chlorobenzoyloxy)-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide
2xcex1-(2xe2x80x2-methoxybenzoyloxy)-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide
2xcex1-(3xe2x80x2-ethylbenzoyloxy)-5,7,8xcex1H-eudesma-4(15),11 (13)-dien-8,12-olide
2xcex1-(4xe2x80x2-ethylbenzoyloxy)-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide
2xcex1-(4xe2x80x2-isopropylbenzoyloxy)-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide
2xcex1-(4xe2x80x2-chlorobenzoyloxy)-5,7,8xcex1H-eudesma-4(15),11 (13)-dien-8,12-olide
2xcex1-(3xe2x80x2,5xe2x80x2-tert-dibutyl-4xe2x80x2-hydroxybenzoyloxy)-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12olide
2xcex1-(2xe2x80x2-methoxy-5xe2x80x2-chlorobenzoyloxy)-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide
2xcex1-(3xe2x80x2-fluoro-4xe2x80x2-methoxybenzoyloxy)-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide
2xcex1-(2xe2x80x2-trifluoromethylbenzoyloxy)-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide
2xcex1-(4xe2x80x2-trifluoromethylbenzoyloxy)-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide
2xcex1-benzoyloxy4,5,7,8,11xcex1H-eudesman-8,12-olide
2xcex1-benzoyloxy-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide
2xcex1-benzoyloxy-5,7,8,11xcex1H-eudesm-4(15)-en-8,12-olide
2xcex2-benzoyloxy-4,5,7,8,11xcex1H-eudesman-8,12-olide
2xcex2-benzoyloxy-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide
2xcex2-benzoyloxy-5,7,8,11xcex1H-eudesm-4(15)-en-8,12-olide
2xcex1-cyclopropanoyloxy-4,5,7,8,11xcex1H-eudesman-8,12-olide
2xcex1-cyclopropanoyloxy-5,7,8,11xcex1H-eudesm-4(15)-en-8,12-olide
2xcex1-cyclopropanoyloxy-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide
2xcex2-cyclopropanoyloxy-4,5,7,8,11xcex1H-eudesman-8,12-olide
2xcex2-cyclopropanoyloxy-5,7,8,11xcex1H-eudesm-4(15)-en-8,12-olide
2xcex2-cyclopropanoyloxy-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide
2xcex1-furoyloxy-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide
2xcex1-furoyloxy-5,7,8,11xcex1H-eudesm-4(15)-en-8,12-olide
2xcex1-furoyloxy-4,5,7,8,11xcex1H-eudesman-8,12-olide
2xcex2-furoyloxy-5,7,8xcex1H-eudesma-4( 15),11(13)-dien-8,12-olide
2xcex2-furoyloxy-5,7,8,11xcex1H-eudesm-4(15)-en-8,12-olide
2xcex2-furoyloxy-4,5,7,8,11xcex1H-eudesman-8,12-olide
2xcex1-isobutyryloxy-5,7,8,11xcex1H-eudesm-4(15)-en-8,12-olide
2xcex1-isobutyryloxy-4,5,7,8,11xcex1H-eudesman-8,12-olide
2xcex1-isobutyryloxy-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide
2xcex2-isobutyryloxy-5,7,8,11xcex1H-eudesm-4(15)-en-8,12-olide
2xcex2-isobutyryloxy-4,5,7,8,11xcex1H-eudesman-8,12-olide
2xcex2-isobutyryloxy-5,7,8xcex1H-eudesma-4( 15),11(13)-dien-8,12-olide
2xcex1-propanoyloxy4,5,7,8,11xcex1H-eudesman-8,12-olide
2xcex1-propanoyloxy-5,7,8,11xcex1H-eudesm-4(15)-en-8,12-olide
2xcex1-propanoyloxy-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide
2xcex2-propanoyloxy-4,5,7,8,11xcex1H-eudesman-8,12-olide
2xcex2-propanoyloxy-5,7,8,11xcex1H-eudesm-4(15)-en-8,12-olide
2xcex2-propanoyloxy-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide
2xcex1-acetoxy-5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide
2xcex1-acetoxy-5,7,8,11xcex1H-eudesm-4(15)-en-8,12-olide
2xcex1-acetoxy-4,5,7,8,11xcex1H-eudesman-8,12-olide
2xcex2-acetoxy-5,7,8xcex1H-eudesma4(15),11(13)-dien-8,12-olide
2xcex2-acetoxy-5,7,8,11xcex1H-eudesm-4(15)-en-8,12-olide
2xcex2-acetoxy-4,5,7,8,11xcex1H-eudesman-8,12-olide
4,5,7,8,11xcex1H-eudesman-8,12-olide
4,7,8xcex1H-eudesma-5(6),11 (13)-dien-8,12-olide
5,7,8xcex1H-eudesma-4(15),11(13)-dien-8,12-olide
5,7,8,11xcex1H-eudesm-4(15)-en-8,12-olide
2xcex1-hydroxy-4(5),11(13)eudesmadien-12,8-olide
1-[sodium2xcex1-benzoyloxy-8xcex2-hydroxy-11,13-dihydro-5,7,8,11xcex1H-eudesm-4(13)en- 12-oatyl]-pyrrolidine hydrochloride
sodium8xcex2-hydroxy-4,5,7,8,11xcex1H-eudesman-12-oate
sodium8xcex2-hydroxy-5,7,8xcex1H-eudesma-4(15),11(13)-dien-12-oate
sodium 8xcex2-hydroxy-4,7,8xcex1H-eudesma-5(6),11(13)-dien-12-oate
sodium 8xcex2-hydroxy-5,7,8,11xcex1H-eudesm-4( 15)-en-12-oate
sodium 2xcex1,8xcex2-dihydroxy-2xcex2-4,5,7,8,11xcex1H-eudesman-12-oate
sodium 2xcex1,8xcex2-dihydroxy-4(15),11(13 )-eudesmadien-12-oate
sodium 2xcex1-benzoyl-8xcex2-hydroxy-2xcex2,4,5,7,8,11xcex1H-eudesman-12-oate
8xcex2-hydroxy-4,7,8xcex1H-eudesama-5,11(13)-dien-12-oic acid
8xcex2-hydroxy-5,7,8,11xcex1H-eudesam-4( 15)-en-12-oic acid
8xcex2-hydroxy-4,5,7,8,11xcex1H-eudesman-12-oic acid
2xcex1,8xcex2-dihydroxy-2xcex2-4,5,7,8,11xcex1H-eudesaman-12-oic acid
2xcex1,8xcex2-dihydroxy-4(15)11(13)-eudesamadien-12-oic acid
2xcex1-benzoyl-8xcex2-hydroxy-2xcex2,4,5,7,8,11xcex1H-eudesaman-12-oic acid
1-[2xcex1-hydroxy-11,12-dihydro-5,7,8xcex1H-eudesm-4(15)-en-8,12-olidyl]-pyrrolidine: 
Following synthesis, the compounds of the present invention can be isolated and purified by established methods to yield pharmaceutically acceptable material for administration to mammals.
The compounds for use in the methods of the present invention can be, and are preferably, administered as a medicament, i.e., a pharmaceutical composition. Preferred are oral and intraperitoneal dosage forms of the compounds.
The pharmaceutical compositions used in the methods of this invention for administration to animals and humans comprise the compound of the present invention in combination with a pharmaceutical carrier or excipient. The medicament can be in the form of tablets (including lozenges and granules), dragees, capsules, pills, ampoules or suppositories comprising the compound of the invention.
xe2x80x9cMedicamentxe2x80x9d as used herein means physically discrete coherent portions suitable for medical administration. xe2x80x9cMedicament in dosage unit formxe2x80x9d as used herein means physically discrete coherent units suitable for medical administration, each containing a daily dose or a multiple (up to four times) or a sub-multiple (down to a fortieth) of a daily dose of the active compound of the invention in association with a carrier and/or enclosed within an envelope. Whether the medicament contains a daily dose or, for example, a half, a third or a quarter of a daily dose will depend on whether the medicament is to be administered once or, for example, twice, three times or four times a day, respectively.
Advantageously, the compositions are formulated as dosage units, each unit being adapted to supply a fixed dose of active ingredients. Tablets, coated tablets, capsules, ampoules and suppositories are examples of preferred dosage forms according to the invention. It is only necessary that the active ingredient constitute an effective amount, i.e., such that a suitable effective dosage will be consistent with the dosage form employed in single or multiple unit doses. The exact individual dosages, as well as daily dosages, will, of course, be determined according to standard medical principles under the direction of a physician or veterinarian..
The compounds of the present invention can also be administered as suspensions, solutions and emulsions of the active compound in aqueous or non-aqueous diluents, syrups, granulates or powders.
Diluents that can be used in pharmaceutical compositions (e.g., granulates) containing the active compound adapted to be formed into tablets, dragees, capsules and pills include, without limitation, the following: (a) fillers and extenders, e.g., starch, sugars, mannitol and silicic acid; (b) binding agents, e.g., carboxymethyl cellulose and other cellulose derivatives, alginates, gelatine and polyvinyl pyrrolidone; (c) moisturizing agents, e.g., glycerol; (d) disintegrating agents, e.g., agar-agar, calcium carbonate and sodium bicarbonate, (e) agents for retarding dissolution, e.g., paraffin, (f) resorption accelerators, e.g., quaternary ammonium compounds; (g) surface active agents, e.g., cetyl alcohol, glycerol monostearate; (h) adsorptive carriers, e.g., kaolin and bentonite, (i) lubricants, e.g., talc, calcium and magnesium stearate and solid polyethylene glycols.
The tablets, dragees, capsules and pills comprising the active compound can have the customary coatings, envelopes and protective matrices, which can contain opacifiers. They can be so constituted that they release the active ingredient only or preferably in a particular part of the intestinal tract, possibly over a period of time. The coatings, envelopes and protective matrices can be made, for example, from polymeric substances or waxes. The compounds of the present invention can also be made up in microencapsulated form together with one or several of the above-mentioned diluents. The diluents to be used in pharmaceutical compositions adapted to be formed into suppositories can, for example, be the usual water-soluble diluents, such as polyethylene glycols and fats (e.g., cocoa oil and high esters, e.g., C14-alcohol with C16-fatty acid) or mixtures of these diluents.
The pharmaceutical compositions which are solutions and emulsions can, for example, contain the customary diluents, such as solvents, dissolving agents and emulsifiers. Specific non-limiting examples of such diluents are water, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils, ground nut oil, glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitol or mixtures thereof For parental administration, solutions and suspensions should be sterile, e.g., water or arachis oil and, if appropriate, blood-isotonic. The pharmaceutical compositions which are suspensions can contain the usual diluents, such as liquid diluents, e.g., water, ethyl alcohol, propylene glycol, surface active agents (e.g., ethoxylated isostearyl alcohols, polyoxyethylene sorbitols and sorbitan esters), microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, or mixtures thereof.
The pharmaceutical compositions can also contain coloring agents and preservatives, as well as perfumes and flavoring additions (e.g., peppermint oil and eucalyptus oil), and sweetening agents, (e.g., saccharin and aspartame). The pharmaceutical compositions will generally contain from 0.5 to 90% of the active ingredient by weight of the total composition. In addition to the compounds of the present invention, the pharmaceutical compositions and medicaments can also contain other pharmaceutically active compounds. Any diluent in the medicaments of the present invention can be any of those mentioned above in relation to the pharmaceutical compositions. Such medicaments can include solvents of molecular weight less than 200 as the sole diluent. It is envisaged that the compounds of the present invention will be administered perorally, parenterally (for example, intramuscularly, intraperitoneally, subcutaneously, transdermally or intravenously), rectally or topically preferably orally or parenterally, especially perlingually, or intravenously.
The dosage rate, e.g., 0.5 to 500 mg/kg of body weight, will be a function of the nature and body weight of the human or animal subject to be treated, the individual reaction of this subject to the treatment, type of formulation in which the active ingredient is administered, the mode in which the administration is carried out and the point in the progress of the disease or interval at which it is to be administered. Thus, it can in some case suffice to use less than a minimum dosage rate, while other cases an upper limit must be exceeded to achieve the desired results. Where larger amounts are administered, it can be advisable to divide these into several individual administrations over the course of the day.
While not wishing to be bound by any particular theory, it is believed in one aspect that by inhibiting the activity of hormone-sensitive lipase, the compounds of the present invention intervene in the lipolysis of stored triglycerides and their metabolism into fatty acids and glycerol. As a metabolic sequella, elevated glucose levels can be reduced. The metabolism of fat provides three times as much caloric energy as do carbohydrates or proteins. As such, it is a fuel source whose utilization is limited only by its relative abundance. In the liver, free fatty acids undergo xcex1-oxidation, which consequently inhibits glycolysis by negative feedback mechanisms while simultaneously promoting gluconeogenesis. Additionally, the released glycerol is utilized as a three-carbon substrate in gluconeogenesis thus contributing to an increased level of hepatic glucose output. Under normal circumstances of metabolic regulation, enhanced lipolysis of stored triglycerides in adipose tissue occurs after prolonged periods of fasting, and is quickly suppressed following ingestion of a meal by the actions of insulin. However, in the insulin-resistant state of type II diabetes, this suppression of lipolysis by insulin is lost, even after eating. Under these conditions, the liver remains in a constant state of gluconeogenesis, and glucose transport into muscle tissue is reduced, resulting in overt hyperglycemia.
The anti-lipolytic effect of the compounds of the present invention in adipose tissue results in a reduction of plasma free fatty acids and glycerol concentrations. In turn, a reversal from gluconeogenesis to glycolysis in the liver can be achieved due to the decreased availability of fatty acids as a fuel source, with an eventual lowering of plasma glucose levels as the is once again reestablished as the primary energy source. Lowering of free fatty acids has an additional impact on the function of the pancreas. In the xcex2-cell, elevations of free fatty acids cause an impairment in the insulin secretory mechanisms, as demonstrated by Unger (1995, Diabetes 44:863-870), with compounds demonstrating NEFA lowering properties preventing and restoring proper xcex2-cell functioning. Thus, the compounds of the present invention thus achieve a lowering or normalization of elevated blood glucose and insulin levels, and amelioration of the insulin-resistant state of muscle in addition to their beneficial effect on circulating lipids.
Taken together, the results from the enzyme inhibition studies, the lipolysis assays and animal studies described herein suggest that the compounds of the present invention possess a unique mechanism of action, and are not expected to display the toxic cellular transformation effects observed with the prior art thiazolidinediones. They are thus highly useful and desirable therapeutic agents having utility in a variety of disease states where the lowering of blood glucose levels and/or the inhibition of lipolysis is necessary for the treatment of the patients. Such disease states include diabetes, and especially maturity-onset, or Type II diabetes, insulin resistance associated with dyslipidemia and normoglycemia referred to as Syndrome X, hypertension and atherosclerosis.
The enzyme hormone-sensitive lipase (HSL) is distinguished from other endogenous lipase enzyme systems in that its activity is governed by the action of various steroid and non-steroid hormones and secreted neurotransmitter substances via receptor-mediated second messenger pathways. Examples of such substances include insulin, corticosterone, and norepinephrine. The primary location of hormone-sensitive lipase is in adipose tissue, where it is involved in the coordinated regulation of lipid metabolism. As mentioned above, the activity of this enzyme is responsible for mobilization of stored triglycerides to provide fuel sources in times of fasting. Recent expression of the gene for mammalian hormone-sensitive lipase in a transfected bacterial vector has provided a means for the isolation of pure hormone-sensitive lipase protein. This has lead to the development of an in vitro assay for assessing direct effects of potential therapeutic agents on the activity of the enzyme itself without the encumbrances of other signaling and regulating entities found in a whole cell system.
An appropriate model for evaluating adipose cell function in tissue culture conditions is the transformed NIH-3T3 cell. Incubation of the 3T3 cell line with dexamethasone and 3-isobutyl-1-methylxanthine (IBMX) results in differentiation from a fibroblastic adipoblast into a mature adipocyte capable of performing insulin-regulated lipogenesis and norepinephrine-mediated lipolysis. As such, the inhibition of lipolytic activity can be assessed for potential agents in whole cell and permeabilized conditions. An assay of this type has been used to correlate the effects of potential therapeutic agents assessed in the purified enzyme system in an appropriate, intact biological circumstance.
Under normal metabolic regulatory control, the breakdown of stored triglyceride in the adipose tissue of mammals is tightly regulated by the hormone insulin. When insulin levels are high, such as after its release from the xcex2-cell of the pancreas in response to high circulating blood levels of glucose, the activity of hormone-sensitive lipase is inhibited. As glucose levels decrease, the levels of insulin decrease in a parallel fashion. At some point during an extended fasting period, the levels of insulin are decreased below those required for inhibition of hormone-sensitive lipase. As a consequence, the catalytic activity of the enzyme is enhanced resulting in the release of non-esterified free fatty acids (NEFA), and glycerol from stored triglyceride reservoirs.
Normal, non-diabetic animals can thus be utilized for determination of hormone-sensitive lipase inhibition by administering test compounds prior to the initiation of a fasting period when insulin levels are high. A decrease in the level of plasma NEFA after an extended fasting period of 18 hrs would be indicative of inhibiting the mobilization of triglyceride depots from the adipose sites through the hormone-sensitive lipase pathway.
In Type II diabetes, the insulin regulatory mechanism for lipolysis does not function properly. Due to the state of insulin-resistance, and despite the elevation of insulin levels over the normal range in certain instances, the inhibition of hormone-sensitive lipase activity is severely diminished such that the lipolytic activity on stored triglycerides is enhanced, resulting in highly elevated fasting plasma NEFA levels. In addition, recent experimental evidence suggests that a transient state of insulin-resistance can be achieved in normal, non-diabetic individuals following the direct infusion of NEFA into the circulatory system. This data would support a pathological role for elevated plasma NEFA in Type II diabetes.
At the present time, the exact etiology for the development of adult-onset Type II diabetes is unknown. However, it is correlated with marked obesity since 70% of this patient population can be categorized as being obese under the general criterion set forth by American Diabetes Association, National Institutes of Health guidelines. As such, non-human mammals exhibiting a similar pathophysiological profiles serve as appropriate models for assessing the anti-dyslipidemic and anti-diabetic properties of potential pharmaceutical agents.
Animal models displaying these overall characteristics include the ob/ob and KK/Ay strains of mice, and the Zucker fa/fa (ZDF) rat. The ob/ob mouse begins to develop a diabetic profile at 4 weeks of age. Plasma glucose values increase up until 9-10 weeks, at which time the levels begin to plateau. In addition to being hyperglycemic, the ob/ob mouse displays elevated levels of NEFA, triglyceride production, and an insulin-resistant state. Furthermore, a parallel increase in plasma insulin levels occurs as a consequence of hyperglycemia. As plasma insulin levels peak, an improvement in hyperglycemia is seen. However, the amount of insulin present is not sufficient to completely normalize the insulin-resistant state during an oral glucose tolerance test.
Overall, the KK/Ay is similar to the ob/ob with some differences being noted. The onset of the Type II profile occurs much later, roughly 3-4 months of age, and persists for as long as 1 year. Both mouse models display hyperinsulinemia, hyperglycemia, insulin-resistance and dyslipidemia similar to that seen in the clinical setting.
The ZDF rat is also similar to the ob/ob and KK/Ay mouse in that all the general attributes of the Type II-like profile with one added exception. In contrast to the mouse models, which exhibit severe hyperinsulinemia, the ZDF rat displays a progressive decline in the ability to secrete insulin from the xcex1-cell beginning at 10 weeks of age, resulting in a state of hypoinsulinemia. As such, this condition is similar in profile to a subtype of Type II diabetic patients which fail to respond to certain classes of pharmacological agents, such as the sulfonylureas, and can require insulin therapy.